Diabetic kidney disease (DKD) is by far the most common cause of end stage kidney disease in the US. Novel insights about the evolution of DKD lesions may help identify new targets for treatment. Most of what we currently know about the structural changes of DKD comes from the studies performed by two-dimensional (2D) transmission electron microscopy (TEM); however, complexity of some of the cellular and subcellular structures demands for 3D studies. Here, we propose to apply serial block face scanning EM (SBF-SEM), a relatively novel technique that allows for 3D EM studies, to study DKD in human and mice. We will optimize SBFSEM tissue preparation protocols, will generate 3D models to better understand the spatial relationships between the glomerular structures. Adding quantitative approached to the volumetric data from SBF-SEM will make this much more powerful. We will develop quantitative approaches to study mitochondrial number and fission/fusion and subpodocyte space volume. Accomplishment of these goals may help finding structural evidence of pathogenetic processes that may be amenable to interventions. These could lead to proof of concept studies where interventions altering these structural variables influence outcomes.

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